Prediction of the B cell epitopes of human heparanase protein and determination of their immunogenicity
10.3760/cma.j.issn.0254-5101.2009.03.001
- VernacularTitle:人类肝素酶B细胞表位的预测和免疫原性鉴定
- Author:
Ling DU
;
Huiju WANG
;
Jianmin YANG
;
Yong FANG
;
Zhongsheng ZHAO
;
Han GAO
;
Zaiyuan YE
- Publication Type:Journal Article
- Keywords:
Heparanase;
B cell epitopes;
Epitope prediction;
Immunogenicity
- From:
Chinese Journal of Microbiology and Immunology
2009;29(3):193-197
- CountryChina
- Language:Chinese
-
Abstract:
Objective To predict the secondary structure and the B cell epitopes of human heparanase protein, and to identify its immunogenicity. Methods The flexible regions of secondary structure and the B cell epitopes of human heparanase amino acid sequence were predicted by DNAStar and Bcepred software. The multiple antigenic peptides (MAP) of the epitopes were synthesized in 8-branch form. Rabbits were immunized with the 8-branch MAPs mixed with a universal T-helper epitope human IL-1β peptide (VQGEESNDK, amino acid 163-171 ). The immunogenicity of the synthesized peptides was evaluated by ELISA, Western blot and immunohistochemistry. Results Amino acid 1 -15 ( MAP1), 279-293 (MAP2) and 175-189(MAP3) of large-subunit of human heparanase protein was predicted as the most potential epitopes of human heparanase protein. All the three synthesized MAPs induced high titer of antibodies. ELISA, Western blot and immunohistochemistry analysis showed all the three MAPs could produce high titer serum antibodies, antibodies induced by MAP1 and MAP2 had high specific binding activity , and MAP2 antibody showed the strongest binding activity with liver cancer tissues. Conclusion The large-subunit No. 1-15, 279-293 amino acid of human heparanase protein may be the B cell preponderant epitopes and the strongest immunogenicity may be No. 279-293 peptide, which provided a theoretic basis for the antibody and vaccine development of heparanase subunit peptide.
